No positive surgical margins. One hundred ten (14.9%) patients were excluded from the study for missing data. One hundred forty four (19.5%) patients GDC-0941 chemical structure were not considered as they were submitted to neoadjuvant hormonal therapy. A total of 486 patients were included in the present analysis and were evaluated for all the variables considered (pathologic tumour stage, tumour grade, serum total PSA and CgA, age). None of these patients had previous or

concomitant history of other malignant disease, adrenal incidentalomas, hepatic and/or renal impairment and/or uncontrolled blood hypertension. Similarly, none of the patients were taking drugs known to alter the metabolism and secretion of CgA, such as nitrates and proton pump inhibitors. An informed consent form was obtained from all patients for all the procedures carried out. The investigation

was approved by the local ethical committee. All patients had a biopsy clinically proven T2-T3 N0 M0 prostate adenocarcinoma, as determined by digital rectal examination, transrectal ultrasonography, bone scan, and computed tomography (CT). All patients were submitted to RRP. All RRP specimens were evaluated at our Institute according to routine procedure by the same expert uropathologist. In all patients the tumour stage was assigned selleck chemical according to the 2002 TNM classification [12]. The tumour grade was described at RRP according to the Gleason score grading system [13]. Blood specimens were obtained in all patients in the early morning, after an overnight fast. In all Thymidylate synthase patients a blood sample was collected in the early morning, after an overnight fast for the determination of serum total PSA and CgA. All samples were obtained at least 3 weeks after any prostate manipulation before the surgical procedure. Blood for serum total PSA and CgA assessments was collected in a frozen vial until Ralimetinib in vitro plasma separation. All serum and plasma samples were immediately frozen and stored at -20 C until analysis. ChromograninA was measured with the enzyme-linked immunoabsorbent assay (ELISA-DakoCytomation, Italy) until April 2005 and with the immunoradiometric assay (CGA-RIACT, CIS BIO INTERNATIONAL-France) thereafter. Chromogranin

A ELISA Kit is designed for the quantitative determination of CgA in human plasma (EDTA or heparin). The kit can be used for measuring CgA in the 10 to 500 U/L range. The ELISA kit is a double antibody sandwich assay where samples and conjugates are incubated simultaneously in antibody-coated wells. The imprecision of the assay is less than 9% over the whole measuring range. CGA-RIACT is a solid-phase two site immunoradiometric assay. Two monoclonal antibodies were prepared against sterically remote sites on the CGA molecule. The first one was coated on the solid phase (coated tube), while the second one, was radio-labelled with iodine 125, and used as a tracer. CGA (molecules or fragments) present in the standard or samples to be tested were “”sandwiched”" between the two antibodies.